| 摘要: |
| 为了研究舰船有害气体、噪声、磁场等特殊环境下人脑特异性基因表达的变化情况,构建了一个20周人胎脑cDNA文库,实验从胎脑组织中抽提总mRNA后,经过一系列酶反应后合成cDNA,分级分离柱除去小片段后克隆到λgt10载体,转染宿主菌E.coli. C600 hf1后文库的包装效率为4.6×106 pfu/μg,cDNA平均插入片段大于1.2 kbp. |
| 关键词: 人脑cDNA文库 滴度 |
| DOI： |
| 分类号:R318 |
| 基金项目: |
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| Construction of cDNA Expression Library from 20-Week Human Embryonic Brain |
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| Abstract: |
| To construct a cDNA expression library from 20-week human embryonic brain, 16.6mg total RNA and 192ug poly(A)+ mRNA have been successively obtained from the 20-week human embryonic brain using \"single-step method\" and by chromatography on oligo-(dT) cellulose. With reverse transcriptase (M-MLV), 10μg mRNA was synthesized into 4.4μg blunt cDNA. After the cDNA was ligated to EcoR I adapter and purified by fractionation, 400ng cDNA of about 100bp-8.0kb was collected, of which 50ng cDNA was inserted into λgt11 phage particles. A cDNA library containing 4.6×10 6 recombinants has been constructed. The capacity of this library and the size of cDNA is suitable for futher study. |
| Key words: human embryonic brain cDNA library titre |
